Molecular Pathogenesis of Fanconi Anemia

范可尼贫血的分子发病机制

• 批准号: 9886762
• 负责人: ALAN D. D'ANDREA
• 金额: $ 43.17万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9886762
• 关键词: ATP phosphohydrolase; Acute Myelocytic Leukemia; Affect; Anemia; BRCA2 gene; Binding; Binding Proteins; Bypass; C-terminal; Characteristics; Chromosomal Instability; Collaborations; Complex; Congenital Abnormality; DNA; DNA Crosslinking Agent; DNA Damage; DNA Interstrand Crosslinking; DNA Repair; DNA Repair Pathway; DNA crosslink; Data; Development; Disease; Dysmyelopoietic Syndromes; Elements; Event; Excision; Fanconi Anemia pathway; Fanconi Anemia-BRCA Pathway; Fanconi anemia protein; Fanconi' s Anemia; Genes; Genetic; Genetic Diseases; Genomic Instability; Grant; Hypersensitivity; Inherited; Knock-out; Knockout Mice; Laboratories; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mitomycins; Molecular; Molecular Conformation; Multiprotein Complexes; Mutation; Nonhomologous DNA End Joining; Pancytopenia; Pathogenesis; Pathway interactions; Patients; Phenotype; Play; Polymerase; Population; Predisposition; Process; Proteins; Rare Diseases; Regulation; Resistance; Role; Site; crosslink; cytotoxic; endonuclease; inhibitor/antagonist; knock-down; malignant breast neoplasm; mouse model; neoplastic cell; novel; nuclease; overexpression; p53-binding protein 1; recessive genetic trait; recruit; repaired; replication stress; scaffold

PROJECT SUMMARY Fanconi Anemia (FA) is a rare autosomal recessive genetic disease characterized by congenital defects, bone marrow failure, cancer predisposition, and cellular hypersensitivity to DNA crosslinking agents (ICLs). FA is caused by mutations in one of 23 genes whose protein products collaborate in a pathway required for removing cytotoxic interstrand crosslinks (ICLs) from DNA. The D'Andrea laboratory has identified many of the molecular players and processes in the FA/BRCA pathway. In collaboration with the Soulier laboratory, we recently identified REV7 as the FA gene, FANCV. This was a key finding, since REV7/FANCV is emerging as a critical protein affecting several different DNA repair processes, by engaging with different binding partners through its C-terminal “seatbelt” domain. When the seatbelt of REV7 is bound to SHLD3, it functions upstream in NHEJ. When the seatbelt of REV7 is bound to REV3, it functions as a translesion (TLS) polymerase, Polζ, required for bypassing an unhooked DNA crosslink, generated by the upstream FA proteins. Thus, REV7 deficiency, like the deficiency of other FA proteins, causes sensitivity to Mitomycin C (MMC), an ICL-inducing agent. Through preliminary studies for this grant, we identified novel REV7 interactors by IP-MS. Interestingly, we identified the binding partner, TRIP13, a novel ATPase which flips REV7 into an open conformation and releases REV3 and inactivates the FA/BRCA pathway. We also identified the REV7-binding protein SHLD2/FAM35A. Knockdown of this protein, like REV7 knockdown, disrupts the FA/BRCA pathway. The Specific Aims for the next five years of this grant are 1) to determine the role of the ATPase TRIP13 in the suppression of the FA/BRCA Pathway 2) to determine the role of the REV7/FANCV protein and its binding partner SHLD2/FAM35A in the activation of the FA/BRCA pathway 3) to employ knockout mouse models for REV7/FANCV and TRIP13 to evaluate the regulation of the FA/BRCA pathway.

项目摘要 Fanconi贫血（FA）是一种罕见的常染色体隐性遗传疾病，其特征是先天性缺陷，骨头 骨髓衰竭，癌症易感性和对DNA交联剂（ICL）的细胞超敏反应。 FA是 由23个基因之一的突变引起 从DNA中去除细胞毒性链间交联（ICL）。 D'Andrea实验室已经确定了许多 FA/BRCA途径中的分子玩家和过程。在与更灵魂的实验室合作的情况下，我们 最近将Rev7确定为FA基因FANCV。这是一个关键发现，因为Rev7/Fancv正在出现 通过与不同的结合伙伴接合，影响几种不同的DNA修复过程的关键蛋白 通过其C端“安全带”域。当Rev7的安全带与SHLD3绑定时，它在上游起作用 在nhej。当Rev7的安全带与Rev3绑定时，它可以用作translesion（TLS）聚合酶POLζ， 绕过上游FA蛋白产生的未连接的DNA交联所必需。那，Rev7 像其他FA蛋白的缺乏一样，缺乏症会引起丝裂霉素C（MMC）的敏感性，ICL诱导 代理人。通过该赠款的初步研究，我们通过IP-MS确定了新颖的Rev7交互器。有趣的是， 我们确定了绑定伙伴Trip13，这是一种新颖的ATPase，将Rev7翻转成一个开放构型，并且 释放Rev3并使FA/BRCA途径失活。我们还确定了Rev7结合蛋白 SHLD2/FAM35A。这种蛋白质的敲低，例如Rev7敲低，破坏了FA/BRCA途径。这 本赠款接下来的五年的具体目的是1）确定ATPase Trip13在 FA/BRCA途径的抑制2）确定Rev7/Fancv蛋白的作用及其结合 FA/BRCA途径的激活中的合作伙伴SHLD2/FAM35A 3） REV7/FANCV和TRIP13评估FA/BRCA途径的调节。